Connect public, paid and private patent data with Google Patents Public Datasets

D-Phenylalanyl-L-prolyl-L-arginine aldehyde sulfate and process for the preparation thereof

Download PDF

Info

Publication number
US4399065A
US4399065A US06337288 US33728882A US4399065A US 4399065 A US4399065 A US 4399065A US 06337288 US06337288 US 06337288 US 33728882 A US33728882 A US 33728882A US 4399065 A US4399065 A US 4399065A
Authority
US
Grant status
Grant
Patent type
Prior art keywords
aldehyde
ml
arginine
acid
phenylalanyl
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
Application number
US06337288
Inventor
Sandor Bajusz
Erzsebet Szell nee Hasenohrl
Eva Barabas
Daniel Bagdy
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Patentbureau Danubia
Gyogyszerkutato Intezet
Original Assignee
Patentbureau Danubia
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Grant date

Links

Images

Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K5/00Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06Dipeptides
    • C07K5/06008Dipeptides with the first amino acid being neutral
    • C07K5/06078Dipeptides with the first amino acid being neutral and aromatic or cycloaliphatic
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00Technologies relating to chemical industry
    • Y02P20/50Improvements relating to the production of products other than chlorine, adipic acid, caprolactam, or chlorodifluoromethane, e.g. bulk or fine chemicals or pharmaceuticals
    • Y02P20/55Synthetic design, e.g. reducing the use of auxiliary or protecting groups

Abstract

The invention relates to D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate, highly stable in aqueous solution, and to a process for preparing it from D-phenylalanyl-L-prolyl-L-arginine aldehyde hemisulfate or NG -carboxy derivative containing an acid-sensitive protecting group at its amino terminal, wherein the acid sensitive amino terminal protecting group or optionally the NG -carboxy group is removed with 1 to 12 N sulfuric acid, applied in 1 to 12 equivalent amounts and the resulting free tripeptide aldehyde sulfate is isolated. The D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate of the invention possesses valuable anticoagulant activity.

Description

The invention relates to D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate, highly stable in aqueous solution, and to a process for the preparation thereof.

It is known that heparin, polyanions of related structure (heparinoids), and coumarin derivatives are being mostly applied in anticoagulant therapy at present. It is a common feature of these agents that they fail to induce direct inhibition of the proteolytic reaction, triggering blood clotting. Heparin as a catalyst accelerates the inhibitory action of one of the plasma inhibitors, antithrombin III, on the enzymes of the coagulation process, primarily that of thrombin, while coumarin derivatives are inhibiting the biosynthesis of proteins containing γ-carboxy-glutamic acid (Gla) moieties. There are four proteins of this type which are involved in the blood coagulation process, one of them being prothrombin. Blood coagulation factors having more or less than the normal number of Gla residues, are inactive, and are not participating in the coagulation process. It should be noted, however, that this inhibition covers the synthesis of the entire range of Gla containing proteins, i.e. one of the natural inhibitors of the coagulation process, protein C (or factor XIV), is also synthesized in inactive form in the presence of coumarin derivatives, which is rather disadvantageous. It is also a characteristic feature that heparin is administered primarily in i.v. infusion, as it is practically inactive at oral application, while coumarin derivatives can only be given orally. Consequently, the effect of heparin may be registered rapidly, within a short period of time, while that of coumarin derivatives--being synthesis inhibitors--only after 24 to 36 hours.

Furthermore it is known that there are tripeptide aldehydes which also exhibit anticoagulant activity; however, contrary to the above agents they enter into direct reaction with thrombin, inhibiting its proteolytic reactions even in the absence of antithrombin III. D-phenylalanyl-L-prolyl-L-arginine aldehyde acetate, described in Hungarian Pat. No. 169,870 as well as D-phenylalanyl-L-prolyl-NG -carboxy-L-arginine aldehyde, described in Belgian Pat. No. 880,844 are both potent thrombin inhibitors.

It was observed that the antithrombin potency of the above synthetic arginine-peptide aldehyde salts--especially that of compounds having a free terminal amino group--i.e. D-phenylalanyl-L-prolyl-L-arginine aldehyde acetate and hydrochloride--is varying and rapidly decreasing upon standing in aqueous solution, making therapeutical application impossible. Though the NG -carboxy derivative of the free tripeptide aldehydes, i.e. D-phenylalanyl-L-prolyl-NG -carboxy-L-arginine aldehyde is retaining its activity in aqueous buffer solution for 20 to 24 hours, after several days, however, there is already a significant reduction in potency and after several months there is a loss of original activity even in solid form.

The invention relates to a new salt of D-phenylalanyl-L-prolyl-L-arginine aldehyde which, contrary to hitherto known products, is stable also in aqueous solution, and to a process for the preparation thereof.

It was found that the stability of diverse salts of D-phenylalanyl-L-prolyl-L-arginine aldehyde was varying in aqueous solution, i.e. isotonic salt solution, to a significant degree. In the course of our tests the peptides were dissolved in concentrations of 10 mg/ml, stored at 5° C., and the ensuing change in antithrombin activity registered for 180 days. The potency was assayed in a system containing the following components:

0.2 ml of 0.5 percent bovine fibrinogen in a 0.9 percent solution of sodium chloride,

0.1 ml of tris(hydroxymethyl)-amino-methane hydrochloride--hydrochloric acid buffer (pH 7.2) containing the peptide solution

0.1 ml of U.S. Standard Human Thrombin (NIH, Bethesda, Md., USA), 10 Unit/ml solution.

The thrombin time of the peptide-free system is 15 s, measured in the "Schnither-Gross Coagulometer".

The activity of the tripeptide aldehyde solution was arbitrarily set up as 100, if the reaction mixture induced a fivefold relative thrombin time at a final concentration of 3.5×10-7 M (in the case of the tripeptide aldehyde sulfate at 0.175 μg/ml).

The test data are summarized in Table I. It is apparent that while the activity of the corresponding hydrochloride in isotonic salt solution is starting to decrease after 5 days, and that of the acetate, citrate, tartrate and tosylate already after several days (similarly to the NG -carboxy derivative of the free tripeptide aldehyde having related properties), the D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate is retaining its antithrombin activity for 90 days. In prolonged stability trials D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate proved to be stable even for 180 days in aqueous medium, and in solid form it similarly failed to lose activity for 6 months.

                                  TABLE 1__________________________________________________________________________Antithrombin activity of tripeptide aldehyde salts in isotonic saltsolution            Relative activity.sup.b after            daysPeptide aldehyde.sup.a            0   5   10  15  20  40  90__________________________________________________________________________D-Phe-Pro-Arg-H.2 CH.sub.3 COOH.sup.cd            70-50                60-40                    40-30                        40-30                            40-30                                35-25                                    30-20D-Phe-Pro-Arg-H.2HCl.sup.c            90-60                90-60                    80-50                        70-40                            60-40                                50-30                                    40-30D-Phe-Pro-Arg/COOH/-H.sup.e            100  80  60  50  30  25  20D-Phe-Pro-Arg-H.H.sub.2 SO.sub.4.sup.f            100 100 100 100 100 100 100__________________________________________________________________________ .sup.a The abbreviations used conform to those established in the literature /i.e. J. Biol. Chem. 247, 977/1972//, Z representing benzyloxycarbonyl, Arg/COOH/and, resp., Arg/Z/N.sup.G --carboxy and N.sup.G --benzyloxycarbonylL-arginine groups, resp./. .sup.b the peptide aldehyde including a fivefold relative thrombin time a a final concentration of 3.5 × 10.sup.-7 M in the reaction mixture has an antithrombin activity of 100. .sup.c Product obtained by the hydrogenolysis of ZD-Phe-Pro-Arg/Z/-H in the presence of one equivalent amount of an acid. .sup.d The antithrombin activity of the corresponding citrate, tartrate and tosylate is changing similarly to that of the acetate. .sup.e Product synthesized according to the process described in Belgian Patent 880 844. .sup.f Product prepared according to the process of the present invention

In order to simulate physiological conditions the antithrombin activity of the tripeptide aldehyde salts as well as that of the carbamic acid derivatives was tested also on human plasma in the following system:

0.2 ml of human citrate plasma,

0.1 ml of tris(hydroxymethyl)-amino-methane hydrochloride--hydrochloric acid buffer solution (pH 7.2) containing the peptide solution, and

0.1 ml of U.S. Standard Human Thrombin (NIH, Bethesda Md., USA), 10 Units/ml solution.

The thrombin time of the peptide-free system is 15 s, measured in the "Schnither-Gross Coagulometer".

              TABLE 2______________________________________Antithrombin activity of tripeptide aldehydederivatives in human plasma             Amount of peptide             /μg/ml.sup.+ / required             to increase throm-             bin time twofold             immediately fol-                           Rela-             lowing dissolu-                           tive             tion of the pep-                           acti-Peptide aldehyde  tide          vity______________________________________D-Phe-Pro-Arg-H.H.sub.2 SO.sub.4             0.020         100D-Phe-Pro-Arg/COOH/-H             0.042          45D-Phe-Pro-Arg-H.2 CH.sub.3 COOH             0.065-0.140   35-14D-Phe-Pro-Arg-H.2 HCl             0.060-0.130   33-15Heparin.sup.a     0.105          19______________________________________ .sup.a Value measured with commercial heparin /132.2  μ/mg, U.S. Ph. XVII/. .sup.+ Amount of the peptide in the reaction mixture.

The data of Table 2 clearly demonstrate that the amount of peptide required to achieve a twofold thrombin time increase compared to the control varies according to the batch used in the case of the acetate and the hydrochloride salts, and is a manifold--in the case of the carbamic acid derivative twofold--of the amount required of the tripeptide aldehyde sulfate.

The in vivo trial of the tripeptide aldehyde derivatives is summarized in Table 3. D-Phenylalanyl-L-prolyl-L-arginine aldehyde sulfate has a significant antithrombin potency in vivo. At intravenous and subcutaneous application its efficacy is in the range of that of heparin, generally applied in therapy, however, it has major advantages compared to it. While heparin, given orally, is inactive, therapeutic effect may be achieved with oral doses of 25 mg/kg of the tripeptide aldehyde sulfate (but only with 50 mg/kg doses of the carbamic acid derivative).

                                  TABLE 3__________________________________________________________________________In vivo trialsDose required for therapeutic effect.sup.a         mg/kg/hour                 mg/kg         i.v. infusion                 s.c.    p.o.Peptide aldehyde         rabbit             dog rabbit                     dog rabbit                             dog__________________________________________________________________________D-Phe-Pro-Arg-H.         --  --  --  --  100 100.2 CH.sub.3 COOHD-Phe-Pro-Arg-H.         --  --  --  --  --  100.2 HClD-Phe-Pro-Arg/COOH/-H         --  3.0 10.0                     6.0 50  50D-Phe-Pro-Arg-H.         1.0 0.5 6.0 6.0 25  25.H.sub.2 SO.sub.4Heparin       0.6 0.5 5.0 2.0 --  --__________________________________________________________________________ .sup.a The therapeutic effect is characterized by the dose required to prolong thrombin time in the whole blood 1.5 to 2.5 fold [Nies, A. S. /1978/ in Clinical Pharmacology /Melmon, K. L. and Morrelli, F. F. Eds./2nd Ed. pp. 303 to 306, Macmillan Publ. Co. Inc. New York; and Versraete, M. and Verwilghen, R./1980/ in Drug Treatment, Principles and Practice of Clinical Pharmacology and Therapeutics, 2nd Ed., Avery G. S. Ed. /1980/ pp. 889 to 952, Edinburgh and London].

The toxicity data of D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate are also more favourable than those of either the acetate or the carbamic acid derivative. The acute toxicity data are summarized in Table 4; this amounted in the case of the tripeptide aldehyde sulfate at oral administration to 2 g/kg.

              TABLE 4______________________________________Acute toxicity data in mice          LD.sub.50 mg/kg            i.v.Peptide aldehyde bolus   i.p.   s.c.  p.o.______________________________________D-Phe-Pro-Arg-H.2CH.sub.3 COOH             9       38    --     960D-Phe-Pro-Arg/COOH/-H.sup.a            --      --     --    1200D-Phe-Pro-Arg-H.H.sub.2 SO.sub.4             45.sup.b                    230    1800  >2000Heparin          no literature data            available______________________________________ .sup.a Due to poor solubility the toxicity data obtained at applications other than p.o. are rather uncertain. .sup.b At intravenous infusion the LD.sub.50 amounted to 58 mg/kg in the rabbit.

Considering the low toxicity and high potency of D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate, the therapeutic index, including both, and being the most characteristic indicator of the therapeutical value of a drug, is more favourable than those of the other tripeptide aldehyde derivatives.

On the basis of intravenous infusion trials in dogs the dose of the human intravenous infusion was established as 1-2 mg/kg/hour.

It was found that D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate may be prepared by a method known per se, by submitting benzyloxycarbonyl-D-phenylalanyl-L-prolyl-NG -benzyloxycarbonyl-L-arginine aldehyde (Z-D-Phe-Pro-Arg)Z)-H) to hydrogenolysis in the presence of equivalent amounts of sulfuric acid. Furthermore it may be simply prepared in sufficient purity from its acid sensitive derivative containing triphenylmethyl or t-alkyloxycarbonyl protective group, i.e. from t-butyloxycarbonyl-D-phenylalanyl-L-prolyl-L-arginine aldehyde hemisulfate or t-butyloxycarbonyl-D-phenylalanyl-L-prolyl-NG -carboxy-L-arginine aldehyde with 1 to 12 N sulfuric acid. At the same time the following methods, known per se, failed to furnish satisfactory results:

a. Acidolysis of the t-butyloxycarbonyl group with sulfuric acid dissolved in acetic acid [Beyerman et al.: in Peptides, 1970 (Ed.: H. Nesvadba), p. 138., North Holland, Amsterdam, 1973].

b. Direct conversion of the D-Phe-Pro-Arg/COOH/-H carbamic acid derivative of Belgian Pat. No. 880,844 with sulfuric acid into the unprotected tripeptide aldehyde sulfate.

c. Transformation of diverse other unprotected tripeptide aldehyde salts, i.e. of D-phenylalanyl-L-prolyl-L-arginine aldehyde acetate of Hungarian Pat. No. 169,870 into its sulfate either with an ion-exchange resin or with sulfuric acid.

The products obtained with either of the a-c methods proved to be of poor homogeneity and/or stability in aqueous solution.

Based on the above the invention relates to D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate and to a process for preparing it from D-phenylalanyl-L-prolyl-L-arginine aldehyde hemisulfate or D-phenylalanyl-L-prolyl-NG -carboxy-L-arginine aldehyde, containing an acid sensitive protecting group at its amino terminal, in which the acid sensitive amino terminal protecting group or optionally the NG -carboxy group is removed with 1 to 12 N sulfuric acid, applied in 1 to 12 equivalent amounts, and the resulting free tripeptide aldehyde sulfate is isolated.

According to a preferred process of the invention the L-arginine lactam, protected at its guanidino group with a benzyloxycarbonyl group, is condensed with t-butyloxycarbonyl-D-phenylalanyl-L-proline, the resulting blocked tripeptide lactam is reduced, and the benzyloxycarbonyl group at the guanidino group of the protected tripeptide aldehyde obtained is submitted to hydrogenolysis in ethanol or tetrahydrofuran containing 30 to 40 percent of water, in the presence of an equivalent amount of sulfuric acid. The resulting tripeptide aldehyde hemisulfate, still protected at its amino terminal, is dissolved in 8 to 12 equivalents, preferably 10 equivalents of 4 to 6 N, preferably 5 N sulfuric acid, and heated for 20 to 40 minutes, preferably 30 minutes to 40° to 60° C., preferably to 50° C., the solution is subsequently neutralized with calcium carbonate, filtered, and preferably freeze-dried.

The product prepared in this way may eventually contain 4 to 6 percent of calcium sulfate, which however does not affect either its biological activity or its therapeutic application.

The invention is further illustrated by but not limited to the following Examples.

The RF values in the Examples are determined by silica gel thin-layer chromatography (Kieselgel G, Reanal, Budapest) in the following systems:

1. Ethyl acetate-pyridine-acetic acid-water--480:20:6:11

2. Ethyl acetate-pyridine-acetic acid-water--60:20:6:11

3. Ethyl acetate-pyridine-acetic acid-water--30:20:6:11.

EXAMPLE 1 D-Phenylalanyl-L-prolyl-L-arginine aldehyde sulfate

t-Butyloxycarbonyl-D-phenylalanyl-L-prolyl-L-arginine aldehyde hemisulfate (2.74 g, 5 mmoles) is dissolved in water (5 ml), 10 N sulfuric acid (5 ml) added at constant stirring, and the mixture heated to 50° C. The solution is stirred for 15 minutes at 50° C., then diluted with ice water (25 ml), and its pH adjusted to 6.5 with calcium carbonate (about 2.25 g) at ice cooling. The precipitated calcium sulfate is filtered, and washed twice with water (5 ml). The filtrate is extracted twice with N butanol (10 ml), concentrated to about 30 ml, filtered, if necessary, and freeze dried. Yield 2.25 g (79 percent) of the title product, containing 4.8 percent of calcium sulfate.

RF =0.35 to 0.40

[α]D 20 =-117±1° (c=1, water).

Analysis calculated for C20 H30 O3 N6.H2 SO4.3 H2 O.0.2 CaSO4 (565.85): Calculated: C, 42.45; H, 6.77; N, 14.85; SO4, 20.37; Ca, 1.41; H2 O, 9.55 percent. Found: C, 42.2; H, 6.9; N, 14.85; SO4, 19.8; Ca, 1.3; H2 O, 9.75 percent.

The starting materials are synthesized according to the following procedure:

Step 1: t-Butyloxycarbonyl-D-phenylalanyl-L-prolyl-NG -benzyloxycarbonyl-L-arginine lactam

t-Butyloxycarbonyl-NG -benzyloxycarbonyl-L-arginine lactam (8.6 g, 22 mmoles, Belgian Pat. No. 880,844) is suspended in ethyl acetate (20 ml), and at 5° C. and constant stirring a solution of 4 M hydrochloric acid in ethyl acetate (40 ml) added to it. The reaction mixture is stirred for 30 minutes under ice cooling, diluted with cool ethyl acetate (100 ml), the precipitate formed filtered, washed with ethyl acetate, and dried at reduced pressure in an exsiccator over potassium hydroxide. The resulting NG -benzyloxycarbonyl-L-arginine lactam hydrochloride is dissolved in dimethylformamide (20 ml), and at -10° C. triethylamine (6.2 ml, 44 mmoles) added to it. The suspension formed is added to the following mixed anhydride.

t-Butyloxycarbonyl-D-phenylalanyl-L-proline [U. Ludescher and R. Schwyzer: Helv Chim. Acta 55, 2052 (1972)])7.25 g, 20 mmoles) and N-methyl-morpholine (2.22 ml, 20 mmoles) are dissolved in dimethylformamide (20 ml). The solution is cooled to -15° C., chloroformic acid isobutyl ester (2.64 ml, 20 mmoles) is added to it at stirring and then after 5 minutes the above solution in dimethylformamide is added. The stirring is continued for 1 hour at -15° C., and for 1 hour at 0° C., then the reaction mixture is diluted with benzene (30 ml), the precipitated salts are filtered and washed twice with benzene (10 ml). The solution of benzene-dimethylformamide is diluted with water (50 ml) and the phases are separated. The aqueous layer is extracted twice with benzene (10 ml), then the combined benzene extracts are washed three times with a solution of 10 percent sodium carbonate (30 ml), water (30 ml), three times with 0.5 N sulfuric acid (30 ml), twice with water (30 ml), and the solution evaporated at reduced pressure following drying over anhydrous sodium sulfate. The evaporation residue is homogenized with petroleum ether, filtered, washed with petroleum ether and air-dried. Yield: 9.65 g (76 percent) of the title product.

RF =0.81 to 0.89.

Step 2: t-Butyloxycarbonyl-D-phenylalanyl-L-prolyl-NG -benzyloxycarbonyl-L-arginine aldehyde

The tripeptide lactam (9.52 g, 15 mmoles, Step 1) is dissolved in tetrahydrofuran (45 ml), and at -20° C. and vigorous stirring lithium aluminium hydride (11.25 mmoles) added to it in tetrahydrofuran (about 28 ml of a 0.4 M solution). The proceeding of the reduction is controlled by thin-layer chromatography [RF 1 =0.71 to 0.77 (lactam) and RF 1 =0.31 to 0.39 (aldehyde)]. If necessary, a further portion of the hydride solution is added. When the reaction is concluded the tetrahydrofuran solution is cautiously acidified with 0.5 N sulfuric acid to pH 3, then diluted in such a way that no precipitation occurs (about 100 ml). The aqueous tetrahydrofuran solution is extracted three times with methylene chloride (75 ml), and the combined methylene chloride extracts washed three times with a solution of 10 percent sodium carbonate (10 ml), then twice with water (10 ml). The methylene chloride solution is subsequently dried over anhydrous sodium sulfate and evaporated at reduced pressure. The evaporation residue is dissolved in benzene (50 ml) and the solution repeatedly evaporated at reduced pressure. Then the dissolving and evaporating is repeated once more. The evaporation residue is worked up with ether, filtered, washed with diethyl ether and air-dried. Yield: 6.9 g (72 percent) of the title product.

RF 1 =0.3 to 0.4.

Step 3: t-Butyloxycarbonyl-D-phenylalanyl-L-prolyl-L-arginine aldehyde hemisulfate

Protected tripeptide aldehyde (6.4 g, 10 mmoles, Step 2) is dissolved in a mixture of water (50 ml), tetrahydrofuran (50 ml) and 1 N sulfuric acid (10 ml) and submitted to hydrogenolysis in the presence of a 10 percent palladium charcoal catalyst (1 g). The proceeding of the reaction is controlled by thin-layer chromatography (RF 2 =0.95 to 1.0 [tripeptide aldehyde protected at its guanidino group) and RF 2 =0.45 to 0.54 (unprotected tripeptide aldehyde)]. After the reaction is concluded the catalyst is filtered, washed with a 50 percent aqueous tetrahydrofuran solution (30 ml), and the combined filtrates concentrated at reduced pressure to about 60 ml. The residue is extracted four times with n-butanol. The n-butanol layers are combined and evaporated at reduced pressure to dryness. The evaporation residue is worked up with a mixture of diethyl ether-diisopropyl ether (1:1), filtered, washed with the above mixture, then dried at reduced pressure in an exsiccator. Yield: 4.4 g (80 percent) of the title compound.

RF 2 =0.45 to 0.54.

[α]D 20 =-65±1° (c=1, water).

Analysis calculated for C25 H38 O5 N6.0.5 H2 SO4 (551.64): Calculated: C, 54.43; H, 7.13; N, 15.23; SO4, 8.71 percent. Found: C, 54.5; H, 7.3; N, 15.2; SO4, 8.7 percent.

EXAMPLE 2 D-Phenylalanyl-L-prolyl-L-arginine aldehyde sulfate

t-Butyloxycarbonyl-D-phenylalanyl-L-prolyl-NG -carboxy-L-arginine aldehyde (2.85 g, 5 mmole) is dissolved in water (5 ml), 10 N sulfuric acid (5 ml) added to it and heated to 50° C. The solution is stirred for 15 minutes at 50° C., then diluted with ice water (25 ml), and its pH adjusted to 6.5 with solid calcium hydroxide (about 1.6 grams) under ice cooling. The precipitated calcium sulfate is filtered and washed twice with water (5 ml). The filtrate is extracted twice with n-butanol (10 ml), concentrated at reduced pressure to about 30 ml, filtered, if necessary, and then freeze-dried. Yield: 2.3 g (81 percent) of the title compound, containing 4.9 percent of calcium sulfate.

RF 3 =0.35 to 0.40.

[α]D 20 =-117±1° (c=1, water).

The starting material is prepared according to the following procedure:

t-Butyloxycarbonyl-D-phenylalanyl-L-prolyl-NG -benzyloxycarbonyl-L-arginine aldehyde (6.4 g, 10 mmoles, Example 1, Step 2) is dissolved in 75 percent aqueous ethanol (100 ml), and submitted to hydrogenolysis in the presence of a 10 percent palladium charcoal catalyst (1 g). The progress of the reaction is controlled by thin-layer chromatography [RF 2 =0.90 to 0.95 (protected tripeptide aldehyde) and 0.45 to 0.55 (NG -carboxy derivative)]. At the end of the reaction the catalyst is filtered, washed with water (30 ml), and the filtrate concentrated to 30-40 ml at reduced pressure. The residue is diluted with water (100 ml), extracted twice with methylene chloride (20 ml), and freeze dried. Yield: 5.1 g (85 percent) of the title product.

RF 2 =0.45 to 0.55.

Amino acid analysis: Phe=0.96; Pro=1 (reference amino acid). M.w. according to the amino acid analysis: 570.

EXAMPLE 3 Preparation of a pharmaceutical composition

The 2-ampoule preparation suitable for 6 or 12 hour intravenous infusion is prepared according to the following:

D-Phenylalanyl-L-prolyl-L-arginine aldehyde sulfate (420-840 mg) and human albumin (40-80 mg) are submitted to joint freeze-drying. The contents of the freeze dried ampoule are dissolved prior to use in sterile, germ-free isotonic salt solution (100-200 ml).

Claims (2)

What we claim is:
1. D-Phenylalanyl-L-prolyl-L-arginine aldehyde sulfate highly stable in aqueous solution.
2. A pharmaceutical composition for use as an anticoagulant which comprises a pharmaceutically acceptable carrier and/or diluent and an effective amount of D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate.
US06337288 1981-01-13 1982-01-05 D-Phenylalanyl-L-prolyl-L-arginine aldehyde sulfate and process for the preparation thereof Expired - Lifetime US4399065A (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
HU7081 1981-01-13
HU70/81 1981-01-13

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US06484888 Continuation-In-Part US4478745A (en) 1981-01-13 1983-04-14 T-Butyloxy-carbonyl-D-phenylalanyl-L-prolyl-L-arginine aldehyde hemisulfate, D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate and process for the preparation thereof

Publications (1)

Publication Number Publication Date
US4399065A true US4399065A (en) 1983-08-16

Family

ID=10947750

Family Applications (2)

Application Number Title Priority Date Filing Date
US06337288 Expired - Lifetime US4399065A (en) 1981-01-13 1982-01-05 D-Phenylalanyl-L-prolyl-L-arginine aldehyde sulfate and process for the preparation thereof
US06484888 Expired - Lifetime US4478745A (en) 1981-01-13 1983-04-14 T-Butyloxy-carbonyl-D-phenylalanyl-L-prolyl-L-arginine aldehyde hemisulfate, D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate and process for the preparation thereof

Family Applications After (1)

Application Number Title Priority Date Filing Date
US06484888 Expired - Lifetime US4478745A (en) 1981-01-13 1983-04-14 T-Butyloxy-carbonyl-D-phenylalanyl-L-prolyl-L-arginine aldehyde hemisulfate, D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate and process for the preparation thereof

Country Status (11)

Country Link
US (2) US4399065A (en)
JP (1) JPH0251920B2 (en)
BE (1) BE891708A (en)
CA (1) CA1182111A (en)
DE (1) DE3200812C2 (en)
DK (1) DK151341C (en)
ES (1) ES8301205A1 (en)
FI (1) FI74024C (en)
FR (1) FR2497799B1 (en)
GB (1) GB2091270B (en)
NL (1) NL8200105A (en)

Cited By (45)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4478745A (en) * 1981-01-13 1984-10-23 Richter Gedeon Vegyeszeti Gyar R.T. T-Butyloxy-carbonyl-D-phenylalanyl-L-prolyl-L-arginine aldehyde hemisulfate, D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate and process for the preparation thereof
DE3505555A1 (en) * 1985-02-18 1986-09-11 Behringwerke Ag New oligopeptidylargininolderivate and their homologous, process for their preparation, their use and containing the same medium
US4703036A (en) * 1984-12-21 1987-10-27 Richter Gedeon Vegyeszeti Gyar Rt Peptide-aldehydes, process for the preparation thereof and pharmaceutical compositions containing the same
US5023236A (en) * 1988-04-07 1991-06-11 Corvas, Inc. Factor VII/VIIA active site inhibitors
EP0526877A2 (en) * 1991-08-06 1993-02-10 Bristol-Myers Squibb Company Peptide aldehydes as antithrombotic agents
WO1993018060A1 (en) * 1992-03-04 1993-09-16 Gyógyszerkutató Intézet Kft. New anticoagulant peptide derivatives and pharmaceutical compositions containing the same as well as a process for the preparation thereof
US5250660A (en) * 1991-11-12 1993-10-05 Eli Lilly And Company Peptide purification process
US5252566A (en) * 1991-11-12 1993-10-12 Eli Lilly And Company Antithrombotic agents
WO1994004651A1 (en) * 1992-08-14 1994-03-03 The Procter & Gamble Company Liquid detergents containing a peptide aldehyde
WO1994017817A1 (en) * 1993-02-12 1994-08-18 Corvas International, Inc. Inhibitors of thrombosis
US5416093A (en) * 1991-11-12 1995-05-16 Eli Lilly And Company Antithrombotic agents
US5430023A (en) * 1990-09-28 1995-07-04 Eli Lilly And Company Tripeptide antithrombotic agents
US5436229A (en) * 1994-03-04 1995-07-25 Eli Lilly And Company Bisulfite adducts of arginine aldehydes
US5439888A (en) * 1994-03-04 1995-08-08 Eli Lilly And Company Antithrombotic agents
US5484772A (en) * 1994-03-04 1996-01-16 Eli Lilly And Company Antithrombotic agents
US5488037A (en) * 1994-03-04 1996-01-30 Eli Lilly And Company Antithrombotic agents
US5576283A (en) * 1992-08-14 1996-11-19 The Procter & Gamble Company Liquid detergents containing a peptide aldehyde
US5578574A (en) * 1994-03-04 1996-11-26 Eli Lilly And Company Antithrombotic agents
US5582762A (en) * 1992-08-14 1996-12-10 The Procter & Gamble Company Liquid detergents containing a peptide trifluoromethyl ketone
US5599793A (en) * 1994-03-04 1997-02-04 Eli Lilly And Company Antithromobotic agents
US5602101A (en) * 1994-03-04 1997-02-11 Eli Lilly And Company Antithrombotic agents
US5656645A (en) * 1994-12-13 1997-08-12 Corvas International, Inc. Aromatic heterocyclic derivatives as enzyme inhibitors
US5658930A (en) * 1994-12-13 1997-08-19 Corvas International, Inc. Aromatic heterocyclic derivatives as enzyme inhibitors
EP0801073A2 (en) 1996-02-12 1997-10-15 Hoechst Aktiengesellschaft Process for the production of oligopeptid-aldehydes
US5705487A (en) * 1994-03-04 1998-01-06 Eli Lilly And Company Antithrombotic agents
US5707966A (en) * 1994-03-04 1998-01-13 Eli Lilly And Company Antithrombotic agents
US5710130A (en) * 1995-02-27 1998-01-20 Eli Lilly And Company Antithrombotic agents
US5721214A (en) * 1995-06-07 1998-02-24 Cor Therapeutics, Inc. Inhibitors of factor Xa
US5726159A (en) * 1994-03-04 1998-03-10 Eli Lilly And Company Antithrombotic agents
US5739354A (en) * 1996-03-26 1998-04-14 Hoechst Marion Roussel, Inc. Process for the preparation of N-methyl-D-phenylalanyl-N- 1- 3- (aminoiminomethyl)amino!propyl!-3,3-difluoro-2-oxohexyl!-L-prolinamide
US5760002A (en) * 1992-12-22 1998-06-02 The Proctor & Gamble Company Diflouro pentapeptide derivative anti-inflammatory agents
US5885967A (en) * 1994-03-04 1999-03-23 Eli Lilly And Company Antithrombotic agents
US5914319A (en) * 1995-02-27 1999-06-22 Eli Lilly And Company Antithrombotic agents
US5919765A (en) * 1995-06-07 1999-07-06 Cor Therapeutics, Inc. Inhibitors of factor XA
US5932733A (en) * 1994-06-17 1999-08-03 Corvas International, Inc. 3-amino-2-oxo-1-piperidineacetic derivatives containing an arginine mimic as enzyme inhibitors
US6022861A (en) * 1995-06-07 2000-02-08 Cor Therapeutics, Inc. Ketoheterocyclic inhibitors of factor Xa
US6046169A (en) * 1995-06-07 2000-04-04 Cor Therapeutics, Inc. Inhibitors of factor XA
US6069130A (en) * 1995-06-07 2000-05-30 Cor Therapeutics, Inc. Ketoheterocyclic inhibitors of factor Xa
US6069232A (en) * 1995-10-02 2000-05-30 Hoechst Marion Roussel, Inc. Polyfluoroalkyl tryptophan tripeptide thrombin inhibitors
US6162783A (en) * 1996-09-24 2000-12-19 The Procter & Gamble Company Liquid detergents containing proteolytic enzyme and protease inhibitors
US6165966A (en) * 1996-09-24 2000-12-26 The Procter & Gamble Company Liquid detergents containing proteolytic enzyme and protease inhibitors
US6180586B1 (en) 1996-09-24 2001-01-30 The Procter & Gamble Company Liquid laundry detergent compositions containing proteolytic enzyme and protease inhibitors
US6245743B1 (en) 1996-06-05 2001-06-12 Cor Therapeutics, Inc. Inhibitors of factor Xa
US20040197162A1 (en) * 1998-11-13 2004-10-07 Pneutools, Incorporated Stacked assembly of roofing caps
US20110028397A1 (en) * 2007-11-30 2011-02-03 Toezser Jozsef Use of Urokinase Type Plasminogen Activator Inhibitors for the Treatment of Corneal Disorders

Families Citing this family (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0118223A1 (en) * 1983-02-07 1984-09-12 Aktiebolaget Hässle Enzyme inhibitors
EP0124317B1 (en) * 1983-04-27 1990-04-11 Imperial Chemical Industries Plc Proline derivatives
US5332726A (en) * 1989-09-29 1994-07-26 Rhone-Poulenc Rorer Pharmaceuticals Inc. Antithrombotic peptides and pseudopeptides
US5064814A (en) * 1990-04-05 1991-11-12 Rhone-Poulenc Rorer Pharmaceuticals Inc. Anti-thrombotic peptide and pseudopeptide derivatives
GB2244994B (en) * 1990-06-12 1994-01-19 Gyogyszerkutato Intezet Improved process for the preparation of tripeptide aldehydes
US5371072A (en) * 1992-10-16 1994-12-06 Corvas International, Inc. Asp-Pro-Arg α-keto-amide enzyme inhibitors
WO1998003540A3 (en) * 1996-07-19 1998-04-09 Menarini Farma Ind 1,2-substituted cycloalkane derivatives as thrombine inhibitors, a process for the preparation thereof and the use thereof in pharmaceutical formulations
EP2343310A1 (en) * 2010-01-08 2011-07-13 Novozymes A/S Serine hydrolase formulation
WO2013004636A1 (en) 2011-07-01 2013-01-10 Novozymes A/S Stabilized subtilisin composition
CN106471110A (en) 2014-07-03 2017-03-01 诺维信公司 Improved stabilization of non-protease enzyme
DE102015208655A1 (en) 2015-05-11 2016-11-17 Henkel Ag & Co. Kgaa enzyme stabilizers
DE102015210828A1 (en) 2015-06-12 2016-12-15 Henkel Ag & Co. Kgaa The phosphate-free liquid dishwashing detergent
DE102016209406A1 (en) 2016-05-31 2017-11-30 Henkel Ag & Co. Kgaa Stabilized enzyme-containing detergents and cleaners

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3826793A (en) * 1968-10-21 1974-07-30 Bofors Ab Anticoagulant peptides related to fibrino peptides
US4316889A (en) * 1979-01-04 1982-02-23 Richter Gedeon Vegyeszeti Gyar Rt. Novel peptidyl-arginine aldehyde derivatives and process for the preparation thereof

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4399065A (en) * 1981-01-13 1983-08-16 Patentbureau Danubia D-Phenylalanyl-L-prolyl-L-arginine aldehyde sulfate and process for the preparation thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3826793A (en) * 1968-10-21 1974-07-30 Bofors Ab Anticoagulant peptides related to fibrino peptides
US4316889A (en) * 1979-01-04 1982-02-23 Richter Gedeon Vegyeszeti Gyar Rt. Novel peptidyl-arginine aldehyde derivatives and process for the preparation thereof

Cited By (53)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4478745A (en) * 1981-01-13 1984-10-23 Richter Gedeon Vegyeszeti Gyar R.T. T-Butyloxy-carbonyl-D-phenylalanyl-L-prolyl-L-arginine aldehyde hemisulfate, D-phenylalanyl-L-prolyl-L-arginine aldehyde sulfate and process for the preparation thereof
US4703036A (en) * 1984-12-21 1987-10-27 Richter Gedeon Vegyeszeti Gyar Rt Peptide-aldehydes, process for the preparation thereof and pharmaceutical compositions containing the same
DE3505555A1 (en) * 1985-02-18 1986-09-11 Behringwerke Ag New oligopeptidylargininolderivate and their homologous, process for their preparation, their use and containing the same medium
US5023236A (en) * 1988-04-07 1991-06-11 Corvas, Inc. Factor VII/VIIA active site inhibitors
US5430023A (en) * 1990-09-28 1995-07-04 Eli Lilly And Company Tripeptide antithrombotic agents
EP0526877A3 (en) * 1991-08-06 1993-04-07 Bristol-Myers Squibb Company Peptide aldehydes as antithrombotic agents
EP0526877A2 (en) * 1991-08-06 1993-02-10 Bristol-Myers Squibb Company Peptide aldehydes as antithrombotic agents
US5380713A (en) * 1991-08-06 1995-01-10 Bristol-Myers Squibb Company Peptide aldehydes as antithrombotic agents
US5250660A (en) * 1991-11-12 1993-10-05 Eli Lilly And Company Peptide purification process
US5252566A (en) * 1991-11-12 1993-10-12 Eli Lilly And Company Antithrombotic agents
US5416093A (en) * 1991-11-12 1995-05-16 Eli Lilly And Company Antithrombotic agents
WO1993018060A1 (en) * 1992-03-04 1993-09-16 Gyógyszerkutató Intézet Kft. New anticoagulant peptide derivatives and pharmaceutical compositions containing the same as well as a process for the preparation thereof
US5576283A (en) * 1992-08-14 1996-11-19 The Procter & Gamble Company Liquid detergents containing a peptide aldehyde
US5840678A (en) * 1992-08-14 1998-11-24 Procter & Gamble Company Liquid detergents containing a peptide trifluoromethyl ketone
US5830840A (en) * 1992-08-14 1998-11-03 The Procter & Gamble Company Liquid detergents containing a peptide aldehyde
WO1994004651A1 (en) * 1992-08-14 1994-03-03 The Procter & Gamble Company Liquid detergents containing a peptide aldehyde
US5582762A (en) * 1992-08-14 1996-12-10 The Procter & Gamble Company Liquid detergents containing a peptide trifluoromethyl ketone
US5760002A (en) * 1992-12-22 1998-06-02 The Proctor & Gamble Company Diflouro pentapeptide derivative anti-inflammatory agents
WO1994017817A1 (en) * 1993-02-12 1994-08-18 Corvas International, Inc. Inhibitors of thrombosis
US5705487A (en) * 1994-03-04 1998-01-06 Eli Lilly And Company Antithrombotic agents
US5484772A (en) * 1994-03-04 1996-01-16 Eli Lilly And Company Antithrombotic agents
US5599793A (en) * 1994-03-04 1997-02-04 Eli Lilly And Company Antithromobotic agents
US5602101A (en) * 1994-03-04 1997-02-11 Eli Lilly And Company Antithrombotic agents
US6124277A (en) * 1994-03-04 2000-09-26 Eli Lilly And Company Antithrombotic agents
US6090787A (en) * 1994-03-04 2000-07-18 Eli Lilly And Company Antithrombotic agents
US5885967A (en) * 1994-03-04 1999-03-23 Eli Lilly And Company Antithrombotic agents
US5439888A (en) * 1994-03-04 1995-08-08 Eli Lilly And Company Antithrombotic agents
US5707966A (en) * 1994-03-04 1998-01-13 Eli Lilly And Company Antithrombotic agents
US5578574A (en) * 1994-03-04 1996-11-26 Eli Lilly And Company Antithrombotic agents
US5436229A (en) * 1994-03-04 1995-07-25 Eli Lilly And Company Bisulfite adducts of arginine aldehydes
US5726159A (en) * 1994-03-04 1998-03-10 Eli Lilly And Company Antithrombotic agents
US5488037A (en) * 1994-03-04 1996-01-30 Eli Lilly And Company Antithrombotic agents
US5932733A (en) * 1994-06-17 1999-08-03 Corvas International, Inc. 3-amino-2-oxo-1-piperidineacetic derivatives containing an arginine mimic as enzyme inhibitors
US5658930A (en) * 1994-12-13 1997-08-19 Corvas International, Inc. Aromatic heterocyclic derivatives as enzyme inhibitors
US5656645A (en) * 1994-12-13 1997-08-12 Corvas International, Inc. Aromatic heterocyclic derivatives as enzyme inhibitors
US5914319A (en) * 1995-02-27 1999-06-22 Eli Lilly And Company Antithrombotic agents
US5710130A (en) * 1995-02-27 1998-01-20 Eli Lilly And Company Antithrombotic agents
US6069130A (en) * 1995-06-07 2000-05-30 Cor Therapeutics, Inc. Ketoheterocyclic inhibitors of factor Xa
US5919765A (en) * 1995-06-07 1999-07-06 Cor Therapeutics, Inc. Inhibitors of factor XA
US6022861A (en) * 1995-06-07 2000-02-08 Cor Therapeutics, Inc. Ketoheterocyclic inhibitors of factor Xa
US6197748B1 (en) 1995-06-07 2001-03-06 Cor Therapeutics, Inc. Ketoheterocyclic inhibitors of factor Xa
US5721214A (en) * 1995-06-07 1998-02-24 Cor Therapeutics, Inc. Inhibitors of factor Xa
US6046169A (en) * 1995-06-07 2000-04-04 Cor Therapeutics, Inc. Inhibitors of factor XA
US6069232A (en) * 1995-10-02 2000-05-30 Hoechst Marion Roussel, Inc. Polyfluoroalkyl tryptophan tripeptide thrombin inhibitors
EP0801073A2 (en) 1996-02-12 1997-10-15 Hoechst Aktiengesellschaft Process for the production of oligopeptid-aldehydes
US5739354A (en) * 1996-03-26 1998-04-14 Hoechst Marion Roussel, Inc. Process for the preparation of N-methyl-D-phenylalanyl-N- 1- 3- (aminoiminomethyl)amino!propyl!-3,3-difluoro-2-oxohexyl!-L-prolinamide
US6245743B1 (en) 1996-06-05 2001-06-12 Cor Therapeutics, Inc. Inhibitors of factor Xa
US6162783A (en) * 1996-09-24 2000-12-19 The Procter & Gamble Company Liquid detergents containing proteolytic enzyme and protease inhibitors
US6180586B1 (en) 1996-09-24 2001-01-30 The Procter & Gamble Company Liquid laundry detergent compositions containing proteolytic enzyme and protease inhibitors
US6165966A (en) * 1996-09-24 2000-12-26 The Procter & Gamble Company Liquid detergents containing proteolytic enzyme and protease inhibitors
US20040197162A1 (en) * 1998-11-13 2004-10-07 Pneutools, Incorporated Stacked assembly of roofing caps
US20060251497A1 (en) * 1998-11-13 2006-11-09 Pneutools, Incorporated Method of forming a stacked assembly of roofing caps
US20110028397A1 (en) * 2007-11-30 2011-02-03 Toezser Jozsef Use of Urokinase Type Plasminogen Activator Inhibitors for the Treatment of Corneal Disorders

Also Published As

Publication number Publication date Type
FI820086A (en) application
DK151341B (en) 1987-11-23 grant
DE3200812C2 (en) 1986-09-18 grant
GB2091270B (en) 1984-08-22 grant
GB2091270A (en) 1982-07-28 application
BE891708A (en) 1982-07-07 grant
ES508637D0 (en) grant
ES508637A0 (en) 1982-11-16 application
BE891708A1 (en) grant
CA1182111A1 (en) grant
JPH0251920B2 (en) 1990-11-08 grant
FI74024B (en) 1987-08-31 application
US4478745A (en) 1984-10-23 grant
JPS57181046A (en) 1982-11-08 application
FR2497799B1 (en) 1985-06-28 grant
FI820086L (en) 1982-07-14 grant
DE3200812A1 (en) 1982-08-12 application
JP1630438C (en) grant
DK151341C (en) 1988-05-09 grant
CA1182111A (en) 1985-02-05 grant
DK9382A (en) 1982-07-14 application
ES8301205A1 (en) 1983-02-16 application
NL8200105A (en) 1982-08-02 application
FI74024C (en) 1987-12-10 grant
FR2497799A1 (en) 1982-07-16 application

Similar Documents

Publication Publication Date Title
US5492895A (en) Inhibitors of thrombosis
US4996358A (en) Hydroxylamine bearing amino acid derivatives as collagenase inhibitors
US4879313A (en) Novel platelet-aggregation inhibitors
US4587046A (en) Drug-carrier conjugates
US4024248A (en) Peptides having LH-RH/FSH-RH activity
US5554728A (en) Lipid conjugates of therapeutic peptides and protease inhibitors
US4732890A (en) Retro-inverso hexapeptide neurotensin analogs, process for their preparation and pharmaceutical compositions containing them
US4346078A (en) Novel anticoagulant agmatine derivatives and process for the preparation thereof
US4910296A (en) Medicaments containing alpha 1 thymosin fragments and having an immunostimulant action, and fragments of alpha 1 thymosin
US6057297A (en) Inhibitor compounds of zinc-dependent metalloproteinases associated with pathological conditions, and therapeutic use thereof
EP0460679A2 (en) Endothelin antagonistic peptide derivatives
US4992463A (en) Thienyl peptide mimetic compounds which are useful in inhibiting platelet aggregation
US5449662A (en) Atrial natriuretic peptide clearance inhibitors which resist degradation
US5420109A (en) Cytokine restraining agents
US5610140A (en) Bradykinin receptor antagonists with neurokinin receptor blocking activity
US5229366A (en) Peptide-containing polyethylene glycol derivatives and application thereof
EP0406428A1 (en) Peptide derivatives and their use
US4737487A (en) VIP type peptides
US4388304A (en) Angiotensin-II analogues with antagonizing effects, containing an ester group in position 8, and a process for the preparation thereof
US4118483A (en) Peptides having gonadoliberin activity and process for their manufacture
US5110797A (en) Peptide compound and a pharmaceutically acceptable salt thereof
US5545719A (en) Nerve growth peptides
EP0260118A1 (en) Selective amidination of diamines
WO1993015756A1 (en) Inhibitors of thrombosis
US5672582A (en) Thrombin inhibitors

Legal Events

Date Code Title Description
AS Assignment

Owner name: RICHTER GEDEON VEGYESZETI GYAR R.T. BUDAPEST , GYO

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNORS:BAJUSZ, SANDOR;SZELL, ERZSEBET;BARABAS, EVA;AND OTHERS;REEL/FRAME:004196/0869

Effective date: 19820309

CC Certificate of correction
FPAY Fee payment

Year of fee payment: 4

FPAY Fee payment

Year of fee payment: 8

AS Assignment

Owner name: GYOGYSZERKUTATO INTEZET KFT, HUNGARY

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNOR:RICHTER GEDEON VEGYESZETI GYAR R.T.;REEL/FRAME:006088/0337

Effective date: 19920310

FPAY Fee payment

Year of fee payment: 12